Compositions and methods for treatment of parkinson's disease

Abstract

The present invention relates to methods for producing neural cells from progenitor or stem cells by activating both the Wnt1-Lmx1a/Lmx1b and the SHH-FoxA2 signaling pathways by, for example, increasing the biological activity of one or more of Wnt1, Lmx1a, Lmx1b, Otx2 and Pitx3 and one or more of SHH, FoxA2 and Nurr1 in the progenitor or stem cells including embryonic stem cells and iPS cells. Such cells may be used for the treatment of Parkinson's disease. The invention further relates to methods for treating Parkinson's disease by increasing the biological activity of one or more of Wnt1, Lmx1b, Lmx1b, Otx2 and Pitx3 and one or more of SHH, FoxA2 and Nurr1 in the midbrain of a patient. In particular, the biological activity of the proteins can be increased by virtue of a cell penetrating peptide fused to the proteins or by transfecting RNAs encoding the proteins such that the host chromosomal DNAs remain intact.

1 . A method for treating Parkinson's Disease in a patient, said method comprising increasing the level of at least one protein selected from the group consisting of Wnt1, Lmx1a, Lmx1b, Otx2 and Pitx3 and at least one protein selected from the group consisting of SHH, FoxA2 and Nurr1 in the midbrain dopaminergic neurons of said patient, wherein the increased biological activity of said proteins is sufficient to treat Parkinson's Disease. 
     
     
         2 . The method of  claim 1 , wherein said midbrain dopaminergic neurons are located in the substantia nigra A9 region. 
     
     
         3 . The method of  claim 1 , wherein the level of FoxA2, Lmx1a and Otx2 is increased in the midbrain dopaminergic neurons of said patient. 
     
     
         4 . The method of  claim 1 , wherein the level of Nurr1, Pitx3 and Lmx1a is increased in the midbrain dopaminergic neurons of said patient. 
     
     
         5 . The method of  claim 1 , wherein the level of Nurr1, Pitx3, Lmx1a, FoxA2 and Otx2 is increased in the midbrain dopaminergic neurons of said patient. 
     
     
         6 . The method of  claim 1 , wherein said method comprises administering to said patient a vector comprising a polynucleotide encoding at least one of the said proteins, operably linked to a promoter, wherein neurons in the patient take up said vector and express said protein. 
     
     
         7 . The method of  claim 6 , wherein said vector is a viral vector. 
     
     
         8 . The method of  claim 7 , wherein said viral vector is selected from the group consisting of an adenovirus, adeno-associated virus, lentivirus, and retrovirus. 
     
     
         9 . The method of  claim 6 , wherein said vector is administered to the substantia nigra. 
     
     
         10 . The method of  claim 1 , wherein said method comprises administering said proteins to said patient. 
     
     
         11 . The method of  claim 10 , wherein at least one protein is encapsulated. 
     
     
         12 . The method of  claim 10 , wherein at least one protein is chemically or recombinantly linked to a cell penetrating peptide (CPP). 
     
     
         13 . The method of  claim 12 , wherein said cell penetrating peptide is from about 15 to about 25 amino acid residues long. 
     
     
         14 . The method of  claim 12 , wherein said cell penetrating peptide comprises at least three basic amino acids selected from the group consisting of arginine, lysine, histidine and combinations thereof. 
     
     
         15 . (canceled) 
     
     
         16 . The method of  claim 12 , wherein the cell penetrating peptide comprises a HIV-TAT peptide. 
     
     
         17 . The method of  claim 1 , wherein said method further comprises increasing the biological activity of at least one of the proteins selected from the group consisting of En1, En2 and Ngn2 in the midbrain dopaminergic neurons of said patient. 
     
     
         18 . A method for producing a neural cell comprising:
 (a) providing a progenitor cell, and   (b) increasing the level of at least one protein selected from the group consisting of Wnt1, Lmx1a, Lmx1b, Otx2 and Pitx3 and at least one protein selected from the group consisting of SHH, FoxA2 and Nurr1 in said progenitor cell under conditions suitable to produce a neural cell.   
     
     
         19 . The method of  claim 18 , wherein said neural cell is a dopaminergic neuron. 
     
     
         20 . The method of  claim 18 , wherein said neural cell expresses tyrosine hydroxylase. 
     
     
         21 . The method of  claim 18 , wherein said neural cell expresses the dopamine transporter or dopa decarboxylase (DDC). 
     
     
         22 .- 53 . (canceled)